Microfluidic platform for the study of intercellular communication viasoluble factor-cell and cell-cell paracrine signaling

摘要

Diffusion of autocrine and paracrine signaling molecules allows cells to communicate in the absence of physical contact. This chemical-based, long-range communication serves crucial roles in tissue function, activation of the immune system, and other physiological functions. Despite its importance, few in vitro methods to study cell-cell signaling through paracrine factors are available today. Here, we report the design and validation of a microfluidic platform that enables (i) soluble molecule-cell and/or (ii) cell-cell paracrine signaling. In the microfluidic platform, multiple cell populations can be introduced into parallel channels. The channels are separated by arrays of posts allowing diffusion of paracrine molecules between cell populations. A computational analysis was performed to aid design of the microfluidic platform. Specifically, it revealed that channel spacing affects both spatial and temporal distribution of signaling molecules, while the initial concentration of the signaling molecule mainly affects the concentration of the signaling molecules excreted by the cells. To validate the microfluidic platform, a model system composed of the signaling molecule lipopolysaccharide, mouse macrophages, and engineered human embryonic kidneycells was introduced into the platform. Upon diffusion from the firstchannel to the second channel, lipopolysaccharide activates the macrophages which begin toproduce TNF-α. The TNF-α diffuses from the second channel to the third channel tostimulate the kidneycells, which express green fluorescent protein (GFP) in response. Byincreasing the initial lipopolysaccharide concentration an increase in fluorescentresponse was recorded, demonstrating the ability to quantify intercellular communicationbetween 3D cellular constructs using the microfluidic platform reportedhere. Overall, these studies provide a detailed analysis on how concentration of theinitial signaling molecules, spatiotemporal dynamics, and inter-channel spacing affectintercellularcommunication.